Bifidobacterium animalis subsp. Lactis strain WKB148 as well as product and application thereof
A technology of animal bifidobacterium and milk subspecies, applied in the field of microorganisms, to achieve broad application prospects, strong tolerance, and increased content
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experiment example 1
[0090]Experimental example 1. Isolation, screening and identification of Bifidobacterium animalis subsp. lactis strain WKB148
[0091] (1) Isolation and screening of strains
[0092] Collect healthy breast milk samples, put 1g of healthy breast milk samples into 9mL sterile normal saline, fully shake and disperse evenly, dilute sterile normal saline to a suitable gradient, take 100μL and spread evenly on Mopi with 5% (V / V) On the MRS agar plate containing Luoxing lithium salt and 0.05% L-cysteine hydrochloride, after culturing under anaerobic conditions at 37°C for 48-72 hours, select opaque milky white, round and shiny, neat edges, surface Protruding and moist monoclonal colonies were repeatedly streaked and purified on MRS solid medium, and the bifidobacteria strains were obtained after cell morphology and individual morphology observation.
[0093] (2) Morphology and 16S rDNA molecular biology identification of the strain
[0094] The morphological identification includ...
experiment example 2
[0103] Experimental Example 2: Tolerance of Bifidobacterium subsp. lactis WKB148 to Artificial Gastrointestinal Fluid
[0104] Simulated artificial gastric juice: prepare PBS solution, add 0.3% pepsin, adjust the pH value to 2.5 with 1mol / L HCL, then fully dissolve and filter with a 0.22μm microporous membrane for sterilization.
[0105] Simulated artificial intestinal juice: Prepare PBS solution, add 0.1% trypsin and 0.3% ox gall powder, adjust the pH value to 8.0 with 0.1mol / L NaOH, then fully dissolve and filter and sterilize with a 0.22μm microporous membrane for use.
[0106] Bifidobacterium animalis subsp. lactis strain WKB148 was activated and cultured for 2 generations under anaerobic conditions. Centrifuge the activated Bifidobacterium animalis milk subsp. milk subsp. WKB148 bacteria liquid, discard the supernatant, collect the bacteria, prepare the bacteria suspension and adjust the number of viable bacteria to 10 8 CFU / mL. After centrifuging 1mL of the bacterial s...
experiment example 3
[0112] Experimental example 3, the inhibitory ability of animal Bifidobacterium lactis subsp. lactis strain WKB to pathogenic bacteria
[0113] The antagonistic strains were inoculated at 2% (V / V) into an anaerobic glass tube containing 0.05% L-cysteine hydrochloride MRS liquid medium, and cultured at a constant temperature of 37°C for 12 hours. Inoculate the pathogenic strains of Escherichia coli, Salmonella, Staphylococcus aureus, Shigella flexneri, and Listeria monocytogenes in liquid beef extract peptone medium respectively, and cultivate overnight at 37°C, 250 rpm, and constant temperature shaker , and then prepare the pathogenic bacteria suspension with a concentration of 10 8 CFU / mL. Cool the MRS solid medium to about 55°C, and mix it with the pathogenic bacteria suspension in a certain proportion, so that the number of viable pathogenic bacteria in the system is 10 6 CFU / mL order of magnitude, and then quickly pour it into the plate with the Oxford cup placed in ad...
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